Table 1.
Down regulation of TIM affects DNA replication rate.
| Cell Line (HEK293) | Tetracycline | TIM/Actin Protein Levels (±SEM)a | p Value (t test)c | BrdU Incorporation Rate (±SEM)b | p Value (t test)c |
|---|---|---|---|---|---|
| Control shRNA | − Tet | 0.884 (±0.103) | 100% | ||
| Control shRNA | + Tet | .715 (±0.139) | 0.3850 (vs. −Tet) | 101.60 (±10.70)% | 0.886 (vs. −Tet) |
| Tim shRNA | −Tet | 0.882 (±0.060) | 100% | ||
| Tim shRNA | +Tet | 0.219 (±0.020) | 0.0005 (vs. −Tet) | 86.48 (±3.97)% | 0.014 (vs. −Tet) |
Sub-saturated TIM and actin protein levels were determined by densitometric scanning of western blot signals from three independent cell extract samples (for each cell line and treatment) analyzed on the same film. In-lane TIM/actin levels were calculated, normalized to the single highest value and averaged.
Incorporation rates were determined by comparing the slope of least squares regression lines drawn through data taken at four time points (each trial included 6 samples at each of four time points [3, 6, 9, 12 hrs]). The +Tet slope values from each cell line of each trial were normalized relative to the –Tet condition and these values averaged (n=4).
Two-tailed Student's t test (unpaired).